关键词: 血吸虫肠相关循环抗原, CAA与CCA交叉反应, Mono－Q－HPLC纯化分离

Abstract: 　AIM: To investigate the specificity differences of the 2 major gut associated diagnostic circulating molecules in Schistosoma japonicum infection, CAA and CCA, and to obtain sufficient purified material for setting up a standard series in quantitative determinations. METHODS: Isolation and purification of the two worm fractions from a trichloroacetic acid (TCA) soluble preparation of S.japonicum adult worm antigen (AWAj TCA) by Mono Q anion exchange chromatography were performed and the specific reactivity of the eluted fractions by antigen capture ELISA (specific for CAA or CCA) with reference to affinity purified preparations of S.mansoni CAA and CCA was analysed. RESULTS: By using an ionic strength gradient, CCA was eluted in two major peaks, an unbound fraction CCA 1, and a major bound fraction CCA 2. Two additional minor peaks, CCA 3 and CCA 4, were eluted at higher ionic strengths. CAA was only detected in a bound fraction, partly overlapping with CCA 3. In the CCA 1 and CCA 2 fractions reactivity was only found in the antigen capture ELISA using anti CCA McAbs both for capture and detection. The CAA fraction was predominantly found to be positive in the antigen capture ELISA using anti CAA McAbs both for capture and detection. However, when using combinations of anti CCA and anti CAA McAbs for capture and detection by ELISA this fraction showed some reactivity. CONCLUSION: Two CCA fractions contain molecules which bear at least two CCA epitopes; while the CAA fraction contains molecules which contain at least two CAA epitopes, and one CCA epitope.

Key words: Schistosoma gut associated circulating antigen, CAA and CCA cross reactivity, Mono Q purification